Determination of trovafloxacin in human body fluids by high-performance liquid chromatography

For the quantitative determination of trovafloxacin (a new naphthyridinone antibacterial agent) in serum and urine a simple isocratic HPLC method with fluorimetric detection is described. Serum was deproteinised with a mixtur e of acetonitrile and perchloric acid. The protein-free extract was separat ed on a reversed-phase column (Nucleosil 100-5 C-18) and quantified by mean s of fluorescence (excitation 275 nm, emission 405 nm). The mobile phase co nsisted of a mixture of 250 ml acetonitrile and 750 ml distilled water cont aining 10 mmol/l tetrabutylammonium phosphate. Urine was diluted with 0.25 mol/l phosphoric acid 1:20 (v/v) which was adjusted to pH 3.6 with sodium h ydroxide solution. Diluted urine samples were separated on a cation-exchang e column (Nucleosil 100-5 SA) and also detected by means of fluorescence. T rovafloxacin was sufficiently separated from endogenous compounds. Results of validation are given. The method was applied successfully to a study of healthy volunteers. (C) 1999 Elsevier Science B.V. All rights reserved.